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Figure Lengend Snippet: Plasma SDF ‐1 levels and western blot analysis on cardiac expression 2 days after MI . (A) The plasma SDF ‐1 levels were higher in the G group and LE + G group than in the LE group or in the control group (n = 5 in each group. Data are presented as mean ± SE .). * P < 0.05 vs control, LE group. (B) The upregulation of the expression of SDF ‐1 was observed in the infarct border area in the G group and LE + G group (n = 5 in each group. Data are presented as mean ± SE .) * P < 0.05 vs Sham, control, LE group

Article Snippet: The SDF‐1 was assessed using SDF‐1 antibody (1:250 abcam) using lysates from cardiac tissues on the day 2 after MI.

Techniques: Western Blot, Expressing

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Figure Lengend Snippet: HMGN-H1 interplay regulates the binding of transcription factors to the Rex1 super-enhancer. (A) Loss of HMGNs does not affect the expression of Oct4, Nanog, or Sox2. Shown are average of three biological replicates of mRNA-seq. (B) IGV snapshots showing loss of OCT4, NANOG and SOX2 binding at the Rex1 super-enhancer in DKO ESCs. Shown are two biological replicates. Red and brown stars indicate location of the proximal enhancer and super-enhancer regions, respectively (as in Figure ​Figure2).2). Arrows point to the location of the transcription factors. Select histone marks at these regulatory regions shown at the bottom. (C) Genomic regions selected for histone H1 ChIP-qPCR experiment. For exact location see primer sequence in Materials and Methods. Brown star denotes the super enhancer region (compare to panel A). (D) Relative H1 occupancy in WT and DKO ESCs at the five non-regulatory and three super-enhancer regions of Rex1 shown on panel C. (E) Elevated H1 occupancy at the Rex1 super-enhancer, but not at adjacent nonregulatory chromatin regions, in DKO cells. Average of values obtained in WT cells are normalized to 1.0. *P < 0.05; **P < 0.01; ****P < 0.0001.

Article Snippet: The antibodies used in chromatin immunoprecipitation include: Rabbit polyclonal to Rex1 (Thermo Fisher Scientific PA5-27567), Rabbit polyclonal to Nanog (Active Motif, 61419), Rabbit polyclonal to Oct4 (abcam, ab19857), Rabbit polyclonal to SOX2 (Abcam #ab97959), and Rabbit Polyclonal Anti-CTCF (Millipore Sigma #07-729), Rabbit polyclonal to H3K27ac (abcam, ab4729), Rabbit polyclonal to H3K4me1(abcam, ab8895); Rabbit polyclonal to H3K4me3 (abcam, ab8580); Rabbit polyclonal to H3K9ac (abcam, ab4441), Rabbit monoclonal to H2BK5ac (abcam, ab40886).

Techniques: Binding Assay, Expressing, Sequencing

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Figure Lengend Snippet: HMGN mediated chromatin interactions enhance Rex1 transcription by facilitating transcription factor binding at the Rex1 locus. (A) Loss of HMGNs does not affect CTCF binding at Rex1 locus. Shown are two biological replicates of CTCF occupancy at the Rex1 locus, as determined by ChIP-seq assay. (B) Model depicting HMGN effects on chromatin interactions and Rex1 expression. The Rex1 locus is located within a chromatin loop formed by the interaction of CTCF molecules flanking the upstream and downstream regions of the locus. In WT cells (left side of panel) the binding of NANOG, OCT4 and SOX2 at the super-enhancer located 5′ to the Rex1 gene facilitates interaction with proximal regulatory elements thereby optimizing Rex1 expression. Loss of HMGN (right side) does not affect CTCF binding but increases H1 occupancy, abolishes the binding of transcription factors to the super-enhancer, and reduces both the DNase I sensitivity and the levels of histone modification that mark active chromatin. These epigenetic changes prevent contact between the Rex1 super-enhancer and Rex1 proximal regulatory sites, thereby downregulating Rex1 expression.

Article Snippet: The antibodies used in chromatin immunoprecipitation include: Rabbit polyclonal to Rex1 (Thermo Fisher Scientific PA5-27567), Rabbit polyclonal to Nanog (Active Motif, 61419), Rabbit polyclonal to Oct4 (abcam, ab19857), Rabbit polyclonal to SOX2 (Abcam #ab97959), and Rabbit Polyclonal Anti-CTCF (Millipore Sigma #07-729), Rabbit polyclonal to H3K27ac (abcam, ab4729), Rabbit polyclonal to H3K4me1(abcam, ab8895); Rabbit polyclonal to H3K4me3 (abcam, ab8580); Rabbit polyclonal to H3K9ac (abcam, ab4441), Rabbit monoclonal to H2BK5ac (abcam, ab40886).

Techniques: Binding Assay, ChIP-sequencing, Expressing, Modification

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Figure Lengend Snippet: (A) Immunolabeling of SOX2, PROM1 and NESTIN in GSC lines (GSC-120 and GSC-14). Scale bar, 12 μm. (B) Single-molecule RNA-FISH images indicate localization of MALAT1, PANTR1 , and LINC01563 lncRNA transcripts (red dots) within the nuclei and/or cytoplasm in GSCs. Scale bars, 12 μm. (C) Subcellular fractionation followed by qRT-PCR confirming the localization of PANTR1 and LINC01563 lncRNA transcripts. GAPDH and MALAT1 were used as cytoplasmic and nuclear markers for quality control, respectively. Data are presented as mean values ± SEM (N=3 independent experiments). (D) Kaplan–Meier overall survival prognosis of LINC01563 and PANTR1 .

Article Snippet: Then the cells were incubated with anti-NESTIN (# ab22035, Abcam), anti-SOX2 (#ab 97959, Abcam) and anti-CD133 (#ab19898, Abcam) antibodies (dilution 1:100) overnight at 4°C.

Techniques: Immunolabeling, Fractionation, Quantitative RT-PCR

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Figure Lengend Snippet: (A) Immunolabeling of SOX2, NESTIN and CD133 in GSC lines (GSC-6 and GSC-161). Scale bar, 12 μm. (B) Single-molecule RNA-FISH images indicate localization of MALAT1, PANTR1 , and LINC01563 lncRNA transcripts (red dots) within nuclei and/or cytoplasm in GSC lines (GSC-6 and GSC-161). Scale bars, 12 μm. (C) Subcellular fractionation of GSCs followed by qRT-PCR confirmed the localization of PANTR1 and LINC01563 lncRNA transcripts. GAPDH and MALAT1 were used as cytoplasmic and nuclear markers for quality control, respectively. Data are presented as mean values ± SEM (N=3 independent experiments). (D) No overall survival prognosis was identified for MALAT1 and PANTR1 in both LGG and GBM.

Article Snippet: Then the cells were incubated with anti-NESTIN (# ab22035, Abcam), anti-SOX2 (#ab 97959, Abcam) and anti-CD133 (#ab19898, Abcam) antibodies (dilution 1:100) overnight at 4°C.

Techniques: Immunolabeling, Fractionation, Quantitative RT-PCR